1) Herstellung <ol><li> Um zu beginnen, starten Sie den Neurostar System, melden Sie sich an, und führen Sie die automatisierten Coil-Test. </li><li> Als nächstes auf die entsprechende Patientenakte aus der Liste (alle behandelten Patienten mit dem Neurostar Systems sind in den zentralen Computer eingeloggt) </li><li> Schließlich gewährleisten die Stuhl-Einstellungen sind nicht restriktiv und heben Sie den Stuhl Arm, um es einfach für den Patienten zu betreten und sich setzen. </li></ol> 2) Sitz-und Ausrichten der Patient <ol><li> Wenn der Patient eintrifft, Sitz ihm / ihr bequem in den Sessel. </li><li> Stellen Sie sicher, sowohl für den Patienten und der Techniker einfügen Ohrstöpsel. </li><li> Mit dem Klebestreifen legen den Kopf Positionierung Gurt direkt über dem Patienten Augenbrauen. Achten Sie darauf, den zentralen Punkt oberhalb des Nasion liegt. </li><li> Recline den Stuhl und stellen Sie die Kopfstütze nach Bedarf. </li><li> Leicht befestigen Sie das obere Kopfband, um den Klettverschluss über dem Patienten Nasion. </li><li> Aktivieren Sie die Laser-und sicherzustellen, dass es halbiert die Patienten Nasion. </li><li> Fragen Sie den Gegenstand an einem Punkt an der Wand anstarren und passen Sie die A / P bar bis es die Höhe der Augäpfel ist. </li><li> Stellen Sie sicher, der Patient vollständig zentriert ist und die Velcro an der Seite Kopfpositionierung Riemen wird der Behandlungsstuhl zu sichern. </li><li> Leicht greifen die Seite Kopfpolster nach kranial Stabilität während der Motor Schwelle Bestimmung zu gewährleisten. </li><li> Anschließend die Kopfstütze System-Einstellungen über den Touchscreen-Schnittstelle. </li></ol> 3) Bestimmen Motor Threshold <ol><li> Um festzustellen, Motor Schwelle, bewegen Sie die Spule auf den Kopf des Patienten, so dass die Spule an der Spitze ist und die Senstar Registerkarte über dem oberen Rand des Ohres ausgerichtet. </li><li> Bewegen Sie den A / P-Bar nach vorne, bis es mit der Seite der Spule ausgerichtet ist. </li><li> Richten Sie die Mittellinie auf die Spule mit der 0-Grad-Marke auf der Spule Winkel-Anzeige und bewegen wieder die A / P-Bar. </li><li> Sobald der Kontakt und die Position festgelegt sind, fragen Sie das Thema auf die Anhalterin Position zu erlassen: Arm gebogen 90 Grad im Ellenbogen, statt die Faust locker, Daumen nach außen erweitert. </li><li> Begin Stimulation und konzentrieren sich auf jede Bewegung des Daumens. </li><li> Erhöhen Sie die MT Niveau von 0,68 bis 1,10 bis Daumen oder Finger übereinstimmende Bewegung zu sehen ist. </li><li> Sobald Bewegung festgestellt wird, beginnt die Suche nach dem Motor-Hotspot. Bewegen Sie die Spule um in einem Raster wie Muster, die Anpassung der MT-Ebene als notwendig. Setzen Sie diese Suche, bis Sie einen Speicherort und Leistung Einstellung, die genau 5 Daumen zuckt entlockt von 10 aufeinander folgenden Pulsen zu finden. </li><li> Sobald der Motor Hotspot bestimmt worden ist, bewegen Sie den A / P bar vorne, bis es wieder, deckt sich mit der Seite der Spule. </li><li> Verwenden Sie den Touchscreen, um die A / P-Stellung, Coil Angle, SOA Position und Power Level aufnehmen. </li><li> Schließlich drücken Sie die Found MT-Taste. Wir sind nun bereit für die Behandlung. </li></ol> 4) Ambulante Behandlung <ol><li> Um zu beginnen, bewegen Sie die Spule 5,5 cm anterior der MT Lage. Dadurch wird die Behandlung Standort. </li><li> Sicherstellen, dass die Spule wird die Kontaktaufnahme mit dem Kopf des Patienten und den Patienten bitten, bleiben für die Dauer der Behandlung. </li><li> Die Behandlung Parameter sind in dem Gerät vorinstalliert. Stimulation wird bei 120% Motor Schwelle mit einer Pulsfolge von 10 Hz für 4 Sekunden, durch eine 26 Sekunden Ruhephase folgte erzeugt werden. Die Behandlung wird insgesamt von 37,5 Minuten dauern dies insgesamt 3.000 Impulsen. </li><li> Drücken Sie die Bestätigungstaste Pulse Sequence-Taste und drücken Sie dann auf die Schaltfläche Start. </li><li> Während der Behandlung in den Raum mit dem Patienten zu bleiben und in regelmäßigen Abständen überprüfen, um sicherzustellen, dass die Spule ist die Kontaktaufnahme mit dem Patienten. </li><li> Nach der Impulsfolge beendet ist, bewegen Sie die Spule auf und weg von der Kopf des Patienten und Abkehr der Seite Kopfstütze Pad. </li><li> Heben Sie die Rückenlehne des Stuhls und heben Sie die Armlehne des Sessels. </li><li> Helfen Sie dem Patienten, wie er / sie steht auf. </li><li> Nehmen Sie Ohrstöpsel und berühren Sie den Logout-Button, um die Behandlung zu beenden. </li></ol> 5) repräsentative Ergebnisse TMS ist in der Regel täglich über einen Zeitraum von 4-6 Wochen angewendet werden. Neuere Daten zeigen manche Patienten anfällig für eine symptomatische Rezidiv nach 4-6 Monaten. 8 Wenn dies geschieht, Praktiker Vielleicht möchten Sie Wartungen Meinung nach Wirkung zu verlängern. Diese Behandlung hat sich als wirksam bei ca. 60% der behandelten Patienten. <img src="/files/ftp_upload/2345/2345fig1.jpg" alt="Tabelle 5" />

<ol>
	<li>Pascual-Leone, A., Davey, M., Wassermann, E. M., Rothwell, J., Puri, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Handbook of Transcranial Magnetic Stimulation. , (2002).</li><li>Walsh, V., Pascual-Leone, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Transcranial Magnetic Stimulation: A Neurochronometrics of Mind. , (2005).</li><li>Horvath, J. C., Perez, J., Forrow, L., Fregni, F., Pascual-Leone, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transcranial+Magnetic+Stimulation:+An+Historical+Evaluation+and+Future+Prognosis+of+Therapeutically+Relevant+Ethical+Concerns.">Transcranial Magnetic Stimulation: An Historical Evaluation and Future Prognosis of Therapeutically Relevant Ethical Concerns.</a> , .</li><li>O'Reardon, J. P., Solvason, H. B., Janicak, P. G., Sampson, S., Isenberg, K. E., Nahas, Z., McDonald, W. M., Avery, D., Fitzgerald, P. B., Loo, C., Demitrack, M. A., George, M. S., Sackheim, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficacy+and+safety+of+transcranial+magnetic+stimulation+in+the+acute+treatment+of+major+depression:+a+multisite+randomized+control+trial.">Efficacy and safety of transcranial magnetic stimulation in the acute treatment of major depression: a multisite randomized control trial.</a> Biological Psychiatry. 62, 1208-1216 (2007).</li><li>George, M. S., Lisanby, S. H., Avery, D., McDonald, W. M., Durkalski, V., Pavlicova, M., Anderson, B., Nahas, Z., Bulow, P., Zarkowski, P., Holtzheimer, P. D., Schwartz, T., Sackheim, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Daily+prefrontal+transcranial+magnetic+stimulation+therapy+for+major+depressive+disorder:+a+sham-controlled+randomized+study.">Daily prefrontal transcranial magnetic stimulation therapy for major depressive disorder: a sham-controlled randomized study.</a> Arch Gen Psychiatry. 67, 507-516 (2010).</li><li>Lisanbym, S. H., Husain, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Daily+Left+Prefrontal+Repetitive+Transcranial+Magnetic+Stimulation+(rTMS)+in+the+Acute+Treatment+of+Major+Depression:+Clinical+Predictors+of+Outcome+in+a+Multisite,+Randomized+Controlled+Clinical+Trial.">Daily Left Prefrontal Repetitive Transcranial Magnetic Stimulation (rTMS) in the Acute Treatment of Major Depression: Clinical Predictors of Outcome in a Multisite, Randomized Controlled Clinical Trial.</a> Neuropsychopharmacology. 34, 522-534 (2008).</li><li>Thase, M., Demitrack, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluating+clinical+significance+of+treatment+outcomes+in+studies+of+resistant+major+depression.">Evaluating clinical significance of treatment outcomes in studies of resistant major depression.</a> Biological Psychiatry. 63, 138-138 (2008).</li><li>Demirtas-Tatlidede, A., Mechanic-Hamilton, D., Press, D. Z., Pearlman, C., Stern, W. M., Thall, M., Pascual-Leone, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+open-label,+prospective+study+of+repetitive+transcranial+magnetic+stimulation+(rTMS)+in+the+long-term+treatment+of+refractory+depression:+reproducibility+and+duration+of+the+antidepressant+effect+in+medication-free+patients.">An open-label, prospective study of repetitive transcranial magnetic stimulation (rTMS) in the long-term treatment of refractory depression: reproducibility and duration of the antidepressant effect in medication-free patients.</a> Journal of Clinical Psychiatry. 69, 930-934 (2008).</li></ol>

Die Videoproduktion dieses Artikels wurde von Neuronetics, welche Instrumente in diesen Studien verwendeten produziert gesponsert.

Bei der Betrachtung TMS als potentielle therapeutische Intervention bei Depression, ist es wichtig zu erkennen, die FDA-Protokoll hoch angegeben ist. Um Beton, ist die FDA-Zulassung bis 10 Hz überschwellige Stimulation, die täglich über 4-6 Wochen mit dem Neurostar Gerät bei Patienten, die nicht zufriedenstellende Verbesserung von einem zu erreichen sind begrenzt, aber nicht mehr als eine, angemessene Antidepressiva-Studien während der aktuellen depressiven Folge. An diesem Punkt ist die Nutzung von alternativen Stimulationsmusters, alternative zeitlichen Verlauf, anderes Gerät, oder alternative Patientenpopulation als off-label. Wie die Ergebnisse zur Unterstützung der Wirksamkeit der TMS-Behandlung 5,6,7 erzeugt werden fortfahren, wird neue Daten, aus denen die Möglichkeit einer symptomatischen Rückfalls auftreten 4-6 Monate nach Behandlungsende. Als dieser Publikation gibt es keine zugelassene Erhaltungstherapie Protokolle. Daher müssen Fragen über zusätzliche Behandlung und / oder Wirkung Wartung berücksichtigt werden und genau überwacht werden, indem einzelne Praktizierende.

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<td>Neuronetics Neurostar TMS Device</td>
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<td>SenStar Treatment Link</td>
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<td>Ear Plugs</td>
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<td>Head Pad Sanitary Sheets</td>
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<td>Alignment Head Strap</td>
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the neurostar tms device: conducting the fda approved protocol for treatment of depression

Die Neuronetics Neurostar Transkranielle Magnetstimulation (TMS) System ist eine Klasse II medizinisches Gerät, das kurze Dauer produziert, gepulste Magnetfelder. Diese schnell wechselnden Felder induzieren elektrische Ströme innerhalb lokalisierte, gezielte Regionen der Hirnrinde, die mit verschiedenen physiologischen und funktionellen Veränderungen im Gehirn verbunden sind. 1,2,3 Im Jahr 2007 O&#39;Reardon et al., Unter Verwendung der Neurostar Gerät, die Ergebnisse einer Industrie-gesponserte, Multisite, randomisierte, Schein-Stimulation kontrollierten klinischen Studie, in der 301 Patienten mit Major Depression, die zuvor nicht gelungen war, in Reaktion auf veröffentlichte mindestens eine adäquate Behandlung mit Antidepressiva-Studie unterzog entweder aktiv oder Schein-TMS über dem linken dorsolateralen präfrontalen Kortex (DLPFC). Die Patienten, die Medikamente kostenlos an die Zeit der Studie waren, erhielt TMS fünfmal pro Woche über 4-6 Wochen. 4 Die Ergebnisse zeigten, dass eine Subpopulation der Patienten (diejenigen, die relativ weniger resistent gegen die Medikamente waren, gescheitert nicht mehr als zwei gute pharmakologische Studien) eine statistisch signifikante Verbesserung auf der Montgomery-Asberg Depression Scale (MADRS), der Hamilton Depression Rating zeigte Scale (HAMD) und verschiedene andere Maßnahmen Ergebnis. Im Oktober 2008, unterstützt durch diese und andere ähnliche Ergebnisse 5,6,7, erhalten Neuronetics das erste und einzige Food and Drug Administration (FDA) die Genehmigung für die klinische Behandlung einer bestimmten Form von Medikamenten-refraktären Depression mit einem TMS-Therapie-Gerät ( FDA-Zulassung K061053). In diesem Papier werden wir die angegebenen FDA genehmigt Neurostar Behandlung von Depressionen Protokoll (nur auf ärztliche Verordnung und durch einen lizenzierten Mediziner entweder in einem in-oder ambulant verabreicht werden).

Watch this Scientific Journal Video about The NeuroStar TMS Device: Conducting the FDA Approved Protocol for Treatment of Depression at JoVE.com

The NeuroStar TMS Device: Conducting the FDA Approved Protocol for Treatment of Depression

In diesem Artikel untersuchen wir die Methodik und Erwägungen, die für die FDA genehmigt Behandlung von Depressionen Protokoll über die Neuronetics Neurostar TMS-Gerät.

Die Neurostar TMS Device: Die Durchführung der FDA genehmigten Protokoll für die Behandlung von Depression

The Neuronetics NeuroStar Transcranial Magnetic Stimulation (TMS) System is a class II medical device that produces brief duration, pulsed magnetic ...

the-neurostar-tms-device-conducting-fda-approved-protocol-for

Nanyang Technological University

Research

JoVE Journal

Neuroscience

39.5K Aufrufe.  Beth Israel Deaconess Medical Center. In diesem Artikel untersuchen wir die Methodik und Erwägungen, die für die FDA genehmigt Behandlung von Depressionen Protokoll über die Neuronetics Neurostar TMS-Gerät.

Serie: The NeuroStar TMS Device: Conducting the FDA Approved Protocol for Treatment of Depression

TMS: Using the Theta-Burst Protocol to Explore Mechanism of Plasticity in Individuals with Fragile X Syndrome and Autism

Fragile X Syndrome (FXS), also known as Martin-Bell Syndrome, is a genetic abnormality found on the X chromosome.1,2 Individuals suffering from FXS display abnormalities in the expression of FMR1 - a protein required for typical, healthy neural development.3 Recent data has suggested that the loss of this protein can cause the cortex to be hyperexcitable thereby affecting overall patterns of neural plasticity.4,5
In addition, Fragile X shows a strong comorbidity with autism: in fact, 30% of children with FXS are diagnosed with autism, and 2 - 5% of autistic children suffer from FXS.6
Transcranial Magnetic Stimulation (a non-invasive neurostimulatory and neuromodulatory technique that can transiently or lastingly modulate cortical excitability via the application of localized magnetic field pulses 7,8) represents a unique method of exploring plasticity and the manifestations of FXS within affected individuals. More specifically, Theta-Burst Stimulation (TBS), a specific stimulatory protocol shown to modulate cortical plasticity for a duration up to 30 minutes after stimulation cessation in healthy populations, has already proven an efficacious tool in the exploration of abnormal plasticity.9,10 
Recent studies have shown the effects of TBS last considerably longer in individuals on the autistic spectrum - up to 90 minutes.11 This extended effect-duration suggests an underlying abnormality in the brain's natural plasticity state in autistic individuals - similar to the hyperexcitability induced by Fragile X Syndrome.
In this experiment, utilizing single-pulse motor-evoked potentials (MEPs) as our benchmark, we will explore the effects of both intermittent and continuous TBS on cortical plasticity in individuals suffering from FXS and individuals on the Autistic Spectrum.

In this article, we examine the effects of Theta-Burst TMS stimulation on cortical plasticity in individuals suffering from Fragile X syndrome and individuals on the autistic spectrum.

TMS: Mit dem Theta-Burst-Protokoll zu Mechasnism der Plastizität in Individuen sich mit Fragile X-Syndrom und Autismus

Fragile X Syndrome (FXS), also known as Martin-Bell Syndrome, is a genetic abnormality found on the X chromosome.1,2 Individuals suffering from FXS ...

Forschung

Neurowissenschaften

State-Dependency Effects on TMS:  A Look at Motive Phosphene Behavior

Transcranial magnetic stimulation (TMS) is a non-invasive neurostimulatory and neuromodulatory technique that can transiently or lastingly modulate cortical excitability (either increasing or decreasing it) via the application of localized magnetic field pulses.1,2 Within the field of TMS, the term state dependency refers to the initial, baseline condition of the particular neural region targeted for stimulation. As can be inferred, the effects of TMS can (and do) vary according to this primary susceptibility and responsiveness of the targeted cortical area.3,4,5

In this experiment, we will examine this concept of state dependency through the elicitation and subjective experience of motive phosphenes. Phosphenes are visually perceived flashes of small lights triggered by electromagnetic pulses to the visual cortex. These small lights can assume varied characteristics depending upon which type of visual cortex is being stimulated. In this particular study, we will be targeting motive phosphenes as elicited through the stimulation of V1/V2 and the V5/MT+ complex visual regions.6

In this article, we examine the effects of visually relevant state dependency on TMS induced motive phosphenic presentations.

State-Dependency Auswirkungen auf TMS: Ein Blick auf Motive Phosphene Behavior

Transcranial magnetic stimulation (TMS) is a non-invasive neurostimulatory and neuromodulatory technique that can transiently or lastingly modulate ...

Modeling Neural Immune Signaling of Episodic and Chronic Migraine Using Spreading Depression In Vitro

Migraine and its transformation to chronic migraine are healthcare burdens in need of improved treatment options. We seek to define how neural immune signaling modulates the susceptibility to migraine, modeled in vitro using spreading depression (SD), as a means to develop novel therapeutic targets for episodic and chronic migraine. SD is the likely cause of migraine aura and migraine pain. It is a paroxysmal loss of neuronal function triggered by initially increased neuronal activity, which slowly propagates within susceptible brain regions. Normal brain function is exquisitely sensitive to, and relies on, coincident low-level immune signaling. Thus, neural immune signaling likely affects electrical activity of SD, and therefore migraine. Pain perception studies of SD in whole animals are fraught with difficulties, but whole animals are well suited to examine systems biology aspects of migraine since SD activates trigeminal nociceptive pathways. However, whole animal studies alone cannot be used to decipher the cellular and neural circuit mechanisms of SD. Instead, in vitro preparations where environmental conditions can be controlled are necessary. Here, it is important to recognize limitations of acute slices and distinct advantages of hippocampal slice cultures. Acute brain slices cannot reveal subtle changes in immune signaling since preparing the slices alone triggers: pro-inflammatory changes that last days, epileptiform behavior due to high levels of oxygen tension needed to vitalize the slices, and irreversible cell injury at anoxic slice centers. 
In contrast, we examine immune signaling in mature hippocampal slice cultures since the cultures closely parallel their in vivo counterpart with mature trisynaptic function; show quiescent astrocytes, microglia, and cytokine levels; and SD is easily induced in an unanesthetized preparation. Furthermore, the slices are long-lived and SD can be induced on consecutive days without injury, making this preparation the sole means to-date capable of modeling the neuroimmune consequences of chronic SD, and thus perhaps chronic migraine. We use electrophysiological techniques and non-invasive imaging to measure neuronal cell and circuit functions coincident with SD. Neural immune gene expression variables are measured with qPCR screening, qPCR arrays, and, importantly, use of cDNA preamplification for detection of ultra-low level targets such as interferon-gamma using whole, regional, or specific cell enhanced (via laser dissection microscopy) sampling. Cytokine cascade signaling is further assessed with multiplexed phosphoprotein related targets with gene expression and phosphoprotein changes confirmed via cell-specific immunostaining. Pharmacological and siRNA strategies are used to mimic and modulate SD immune signaling.

Modeling Neural Immune Signaling of Episodic and Chronic Migraine Using Spreading Depression In Vitro

Migraine and its transformation to chronic migraine are immense healthcare burdens in need of improved treatment options. We seek to define how neural immune signaling modulates the susceptibility to migraine, modeled in vitro using spreading depression in hippocampal slice cultures, as a means to develop novel therapeutic targets.

Modeling Neural Immune Signalisierung von episodischen und chronischen Migräne mit Spreading Depression In Vitro</em

Migraine and its transformation to chronic migraine are healthcare burdens in need of improved treatment options.  We seek to define how neural immune ...

Telomerase Activity in the Various Regions of Mouse Brain: Non-Radioactive Telomerase Repeat Amplification Protocol (TRAP) Assay

Telomerase, a ribonucleoprotein, is responsible for maintaining the telomere length and therefore promoting genomic integrity, proliferation, and lifespan. In addition, telomerase protects the mitochondria from oxidative stress and confers resistance to apoptosis, suggesting its possible importance for the surviving of non-mitotic, highly active cells such as neurons. We previously demonstrated the ability of novel telomerase activators to increase telomerase activity and expression in the various mouse brain regions and to protect motor neurons cells from oxidative stress. These results strengthen the notion that telomerase is involved in the protection of neurons from various lesions. To underline the role of telomerase in the brain, we here compare the activity of telomerase in male and female mouse brain and its dependence on age. TRAP assay is a standard method for detecting telomerase activity in various tissues or cell lines. Here we demonstrate the analysis of telomerase activity in three regions of the mouse brain by non-denaturing protein extraction using CHAPS lysis buffer followed by modification of the standard TRAP assay.
In this 2-step assay, endogenous telomerase elongates a specific telomerase substrate (TS primer) by adding TTAGGG 6 bp repeats (telomerase reaction). The telomerase reaction products are amplified by PCR reaction creating a DNA ladder of 6 bp increments. The analysis of the DNA ladder is made by 4.5% high resolution agarose gel electrophoresis followed by staining with highly sensitive nucleic acid stain.
Compared to the traditional TRAP assay that utilize 32P labeled radioactive dCTP&#39;s for DNA detection and polyacrylamide gel electrophoresis for resolving the DNA ladder, this protocol offers a non-toxic time saving TRAP assay for evaluating telomerase activity in the mouse brain, demonstrating the ability to detect differences in telomerase activity in the various female and male mouse brain region.

Telomerase Activity in the Various Regions of Mouse Brain: Non-Radioactive Telomerase Repeat Amplification Protocol TRAP Assay

Telomerase is expressed in the neonatal brain and also in distinct regions of the adult brain. We present a non-toxic time saving TRAP assay for the analysis of telomerase activity in various regions of the mouse brain and detection of differences in telomerase activity between male and female mouse brains.

Telomerase-Aktivität in den verschiedenen Regionen der Maus Gehirn: Nicht-radioaktive Telomerase Repeat Amplification Protocol (TRAP)-Assay

Telomerase, a ribonucleoprotein, is responsible for maintaining the telomere length and therefore promoting genomic integrity, proliferation, and ...

Preparation of Acute Brain Slices Using an Optimized N-Methyl-D-glucamine Protective Recovery Method

This protocol is a practical guide to the N-methyl-D-glucamine (NMDG) protective recovery method of brain slice preparation. Numerous recent studies have validated the utility of this method for enhancing neuronal preservation and overall brain slice viability. The implementation of this technique by early adopters has facilitated detailed investigations into brain function using diverse experimental applications and spanning a wide range of animal ages, brain regions, and cell types. Steps are outlined for carrying out the protective recovery brain slice technique using an optimized NMDG artificial cerebrospinal fluid (aCSF) media formulation and enhanced procedure to reliably obtain healthy brain slices for patch clamp electrophysiology. With this updated approach, a substantial improvement is observed in the speed and reliability of gigaohm seal formation during targeted patch clamp recording experiments while maintaining excellent neuronal preservation, thereby facilitating challenging experimental applications. Representative results are provided from multi-neuron patch clamp recording experiments to assay synaptic connectivity in neocortical brain slices prepared from young adult transgenic mice and mature adult human neurosurgical specimens. Furthermore, the optimized NMDG protective recovery method of brain slicing is compatible with both juvenile and adult animals, thus resolving a limitation of the original methodology. In summary, a single media formulation and brain slicing procedure can be implemented across various species and ages to achieve excellent viability and tissue preservation.

Preparation of Acute Brain Slices Using an Optimized N-Methyl-D-glucamine Protective Recovery Method

This protocol demonstrates the implementation of an optimized N-methyl-D-glucamine (NMDG) protective recovery method of brain slice preparation. A single media formulation is used to reliably obtain healthy brain slices from animals of any age and for diverse experimental applications.

Vorbereitung der akuten Gehirnscheiben mit einem optimierten N-Methyl-D-Glucamine-schützende Recovery-Methode

This protocol is a practical guide to the N-methyl-D-glucamine (NMDG) protective recovery method of brain slice preparation. Numerous recent studies have ...

Receptor Autoradiography Protocol for the Localized Visualization of Angiotensin II Receptors

This protocol describes receptor binding patterns for Angiotensin II (Ang II) in the rat brain using a radioligand specific for Ang II receptors to perform receptor autoradiographic mapping.
Tissue specimens are harvested and stored at -80 &#176;C. A cryostat is used to coronally section the tissue (brain) and thaw-mount the sections onto charged slides. The slide-mounted tissue sections are incubated in 125I-SI-Ang II to radiolabel Ang II receptors. Adjacent slides are separated into two sets: &#39;non-specific binding&#39; (NSP) in the presence of a receptor saturating concentration of non-radiolabeled Ang II, or an AT1 Ang II receptor subtype (AT1R) selective Ang II receptor antagonist, and &#39;total binding&#39; with no AT1R antagonist. A saturating concentration of AT2 Ang II receptor subtype (AT2R) antagonist (PD123319, 10 &#181;M) is also present in the incubation buffer to limit 125I-SI-Ang II binding to the AT1R subtype. During a 30 min pre-incubation at ~22 &#176;C, NSP slides are exposed to 10 &#181;M PD123319 and losartan, while &#39;total binding&#39; slides are exposed to 10 &#181;M PD123319. Slides are then incubated with 125I-SI-Ang II in the presence of PD123319 for &#39;total binding&#39;, and PD123319 and losartan for NSP in assay buffer, followed by several &#39;washes&#39; in buffer, and water to remove salt and non-specifically bound radioligand. The slides are dried using blow-dryers, then exposed to autoradiography film using a specialized film and cassette. The film is developed and the images are scanned into a computer for visual and quantitative densitometry using a proprietary imaging system and a spreadsheet. An additional set of slides are thionin-stained for histological comparisons.
The advantage of using receptor autoradiography is the ability to visualize Ang II receptors in situ, within a section of a tissue specimen, and anatomically identify the region of the tissue by comparing it to an adjacent histological reference section.

Here we present a protocol to describe the localization of angiotensin II Type 1 receptors in the rat brain by quantitative, densitometric, in vitro receptor autoradiography using an iodine-125 labeled analog of angiotensin II.

Rezeptor-Autoradiographie-Protokoll für die Lokalisierte Visualisierung von Angiotensin II-Rezeptoren

This protocol describes receptor binding patterns for Angiotensin II (Ang II) in the rat brain using a radioligand specific for Ang II receptors to ...

Protocol for Isolating the Mouse Circle of Willis

The cerebral arterial circle (circulus arteriosus cerebri) or circle of Willis (CoW) is a circulatory anastomosis surrounding the optic chiasma and hypothalamus that supplies blood to the brain and surrounding structures. It has been implicated in several cerebrovascular disorders, including cerebral amyloid angiopathy (CAA)-associated vasculopathies, intracranial atherosclerosis and intracranial aneurysms. Studies of the molecular mechanisms underlying these diseases for the identification of novel drug targets for their prevention require animal models. Some of these models may be transgenic, whereas others will involve isolation of the cerebro-vasculature, including the CoW.The method described here is suitable for CoW isolation in any mouse lineage and has considerable potential for screening (expression of genes, protein production, posttranslational protein modifications, secretome analysis, etc.) studies on the large vessels of the mouse cerebro-vasculature. It can also be used for ex vivo studies, by adapting the organ bath system developed for isolated mouse olfactory arteries.

We describe here a reproducible protocol for isolating the mouse circle of Willis.

Protokoll für die Maus Kreis von Willis Trenn

The cerebral arterial circle (circulus arteriosus cerebri) or circle of Willis (CoW) is a circulatory anastomosis surrounding the optic chiasma and ...

A Simple One-step Dissection Protocol for Whole-mount Preparation of Adult Drosophila Brains

There is an increasing interest in using Drosophila to model human brain degenerative diseases, map neuronal circuitries in adult brains, and study the molecular and cellular basis of higher brain functions. A whole-mount preparation of adult brains with well-preserved morphology is critical for such whole brain-based studies, but can be technically challenging and time-consuming. This protocol describes an easy-to-learn, one-step dissection approach of an adult fly head in less than 10 s, while keeping the intact brain attached to the rest of the body to facilitate subsequent processing steps. The procedure helps remove most of the eye and tracheal tissues normally associated with the brain that can interfere with the later imaging step, and also places less demand on the quality of the dissecting forceps. Additionally, we describe a simple method that allows convenient flipping of the mounted brain samples on a coverslip, which is important for imaging both sides of the brains with similar signal intensity and quality. As an example of the protocol, we present an analysis of dopaminergic (DA) neurons in adult brains of WT (w1118) flies. The high efficacy of the dissection method makes it particularly useful for large-scale adult brain-based studies in Drosophila.

A Simple One-step Dissection Protocol for Whole-mount Preparation of Adult Drosophila Brains

The adult Drosophila brain is a valuable system for studying neuronal circuitry, higher brain functions, and complex disorders. An efficient method to dissect whole brain tissue from the small fly head will facilitate brain-based studies. Here we describe a simple, one-step dissection protocol of adult brains with well-preserved morphology.

Eine einfache Ein-Schritt-Dissection-Protokoll für Whole-mount Vorbereitung von Adult<em&gt; Drosophila</em&gt; Brains

There is an increasing interest in using Drosophila to model human brain degenerative diseases, map neuronal circuitries in adult brains, and study the ...

A Protocol for the Administration of Real-Time fMRI Neurofeedback Training

Neurologic disorders are characterized by abnormal cellular-, molecular-, and circuit-level functions in the brain. New methods to induce and control neuroplastic processes and correct abnormal function, or even shift functions from damaged tissue to physiologically healthy brain regions, hold the potential to dramatically improve overall health. Of the current neuroplastic interventions in development, neurofeedback training (NFT) from functional Magnetic Resonance Imaging (fMRI) has the advantages of being completely non-invasive, non-pharmacologic, and spatially localized to target brain regions, as well as having no known side effects. Furthermore, NFT techniques, initially developed using fMRI, can often be translated to exercises that can be performed outside of the scanner without the aid of medical professionals or sophisticated medical equipment. In fMRI NFT, the fMRI signal is measured from specific regions of the brain, processed, and presented to the participant in real-time. Through training, self-directed mental processing techniques, that regulate this signal and its underlying neurophysiologic correlates, are developed. FMRI NFT has been used to train volitional control over a wide range of brain regions with implications for several different cognitive, behavioral, and motor systems. Additionally, fMRI NFT has shown promise in a broad range of applications such as the treatment of neurologic disorders and the augmentation of baseline human performance. In this article, we present an fMRI NFT protocol developed at our institution for modulation of both healthy and abnormal brain function, as well as examples of using the method to target both cognitive and auditory regions of the brain.

The ability to induce and/or control neural plasticity may be critical in future treatments for neurologic disorders and the recovery from brain injury. In this paper, we present a protocol on the use of neurofeedback training with functional magnetic resonance imaging to modulate human brain function.

Ein Protokoll für die Verwaltung von Real-Time-fMRT Neurofeedback Training

Neurologic disorders are characterized by abnormal cellular-, molecular-, and circuit-level functions in the brain. New methods to induce and control ...

Using a Microfluidics Device for Mechanical Stimulation and High Resolution Imaging of C. elegans

One central goal of mechanobiology is to understand the reciprocal effect of mechanical stress on proteins and cells. Despite its importance, the influence of mechanical stress on cellular function is still poorly understood. In part, this knowledge gap exists because few tools enable simultaneous deformation of tissue and cells, imaging of cellular activity in live animals, and efficient restriction of motility in otherwise highly mobile model organisms, such as the nematode Caenorhabditis elegans. The small size of C. elegans makes them an excellent match to microfluidics-based research devices, and solutions for immobilization have been presented using microfluidic devices. Although these devices allow for high-resolution imaging, the animal is fully encased in polydimethylsiloxane (PDMS) and glass, limiting physical access for delivery of mechanical force or electrophysiological recordings. Recently, we created a device that integrates pneumatic actuators with a trapping design that is compatible with high-resolution fluorescence microscopy. The actuation channel is separated from the worm-trapping channel by a thin PDMS diaphragm. This diaphragm is deflected into the side of a worm by applying pressure from an external source. The device can target individual mechanosensitive neurons. The activation of these neurons is imaged at high-resolution with genetically-encoded calcium indicators. This article presents the general method using C. elegans strains expressing calcium-sensitive activity indicator (GCaMP6s) in their touch receptor neurons (TRNs). The method, however, is not limited to TRNs nor to calcium sensors as a probe, but can be expanded to other mechanically-sensitive cells or sensors.

Using a Microfluidics Device for Mechanical Stimulation and High Resolution Imaging of C. elegans

New tools for mechanobiology research are needed to understand how mechanical stress activates biochemical pathways and elicits biological responses. Here, we showcase a new method for selective mechanical stimulation of immobilized animals with a microfluidic trap allowing high-resolution imaging of cellular responses.